Shenzhen Key Laboratory of Biochip Research, Biotech and Health Center, City University of Hong Kong Shenzhen Research Institute, Shenzhen 518057, People's Republic of China.
Biosens Bioelectron. 2010 Jul 15;25(11):2402-7. doi: 10.1016/j.bios.2010.02.032. Epub 2010 Mar 6.
In this study, a novel microfluidic device with microbead array was developed and sensitive genotyping of HBV was demonstrated using quantum dot as labels. This device was assembled by using two PDMS slabs featured with different microstructures and channel depths for the construction of a functional region comprising a chamber array and a single sampling microchannel. Since the chamber array and its sampling channel are of different channel depths and are bonded face-to-face, weir structures are generated to confine the microbeads which could be addressed using the microfluidic channel. Highly sensitive virus DNA detection was achieved by the enhanced mass transport in the microfluidics and the rapid reaction dynamics of suspension microbead array. The device could detect 1000 copies/mL of HBV virus in clinical serum samples using in vitro transcribed RNA as the target molecules. Based on DNA hybridization with quantum dots labels, on-chip virus genotyping was also demonstrated with high discrimination specificity and sensitivity (4 pM, S/N >3) using synthesized HBV DNA probes. This microfluidic device combines the rapid binding kinetics of homogeneous assays of microbead array, the liquid handling capability of microfluidics, and the fluorescence detection sensitivity of quantum dots to provide a platform for high sensitivity virus DNA analysis with small reagent consumption, short assay time and parallel detection.
在这项研究中，开发了一种具有微珠阵列的新型微流控装置，并使用量子点作为标记物对 HBV 进行了灵敏的基因分型。该装置由两块具有不同微结构和通道深度的 PDMS 平板组装而成，用于构建一个由腔阵列和单个采样微通道组成的功能区域。由于腔阵列及其采样通道具有不同的通道深度并且面对面粘合，因此会产生堰结构来限制可以使用微流道寻址的微珠。通过微流控中的增强质量传输和悬浮微珠阵列的快速反应动力学，实现了高度灵敏的病毒 DNA 检测。该装置可以使用体外转录 RNA 作为靶分子，从临床血清样本中检测到 1000 拷贝/mL 的 HBV 病毒。基于与量子点标记物的 DNA 杂交，使用合成的 HBV DNA 探针还在芯片上实现了具有高区分特异性和灵敏度（4 pM，S/N >3）的病毒基因分型。这种微流控装置结合了微珠阵列均相分析的快速结合动力学、微流控的液体处理能力以及量子点的荧光检测灵敏度，为具有小试剂消耗、短分析时间和并行检测的高灵敏度病毒 DNA 分析提供了一个平台。
